pT1NX乳酸菌分泌型表达载体 BioVector NTCC质粒载体菌种细胞基因保藏中心

pT1NX乳酸菌分泌型表达载体 BioVector NTCC质粒载体菌种细胞基因保藏中心 Name pT1NX Type Plasmid Class Recombinant Literature WO0023471 Use of a cytokine-producing Lactococcus strain to treat colitis Host_for_distribution Lactococcus lactis subsp. cremoris MG1363 Medium M17 + 0.5% glu (GM17) + erythromycin (5 microg/ml) Selectable_phenotype Erythromycin (ery) Replicon Enterococcus faecalis plasmid pAMbeta1 origin, incl. repD and repE Host_range Lactococcus lactis Gram-positive bacterial strains Cloned_gene Lactococcus usp45 protein gene; secretion signal Staphylococcus aureus protein A gene (spa); cell wall anchor (spaX) Promoter Lactococcus lactis P1 promoter Ribosome_binding_site Ribosome binding site (RBS) of the phage T7 gene 10 (T7g10); modified sequence Terminator Phage T7 gene 10 terminator (T7g10) Further_information The plasmid carries the lactococcal usp45 secretion signal sequence and the sequence encoding the cell wall anchor of S. aureus protein A. pT1NX is an expression vector designed for expression of cloned coding sequences in Gram-positives, particularly lactic acid bacterial (LAB) species, under control of the constitutive P1 promoter. Depending on the insertion site of the coding region, the encoded product is: a) localized in the cytoplasm: when intracellular accumulation is desired, the fragment can be cloned into the unique BglII site. Note that the coding region must then carry a functional ribosome binding site; b) secreted in the medium: secretion of the desired polypeptide is obtained by insertion of the encoding DNA fragment between the unique NaeI and SpeI sites. Insertion at the NaeI site should be done in such a way that the first amino acid of the coding region is joined in frame to the ultimate amino acid of the Usp45 secretion signal; c) displayed at the cell surface: cell wall anchoring and functional surface display of the desired polypeptide is obtained by insertion of the coding region between the unique NaeI and BamHI sites in such a way that an in frame fusion is established with both the Usp45 secretion signal and the protein A anchor. Sequence_detail Sequence at the usp-spaX junction: usp45 --> --> spa anchor 5' ... TAC.GCC.GGCGACGGAT.CCA.AAA ... 3' NaeI BamHI Punctuation indicates reading frame. BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心www.biovector.net [Supplier来源] http://www.biovector.net